Field of the Invention
This present invention relates to compositions and methods for inhibiting nucleic acid polymerases. These compositions and methods may be used for nucleic acid synthesis, amplification, sequencing and cloning.
Related Art
Nucleic acid polymerases (“polymerases”) are enzymes that catalyze the synthesis of nucleic acid molecules that are complementary to a nucleic acid template. Template-directed nucleic acid synthesis is an important aspect of many molecular biology research and diagnostic techniques and assays. Such techniques and assays typically involve extension of a nucleic acid primer designed to hybridize to a specific region of the template.
The yield and homogeneity of primer extension products made by polymerases can be adversely affected by “mispriming,” i.e., hybridization of primers to inappropriate regions of the template, or to non-template nucleic acids. Extension of misprimed nucleic acids can produce high background and obscure detection of properly primed primer extension products. In addition, diversion of nucleic acid synthesis reaction constituents to extend misprimed nucleic acids can reduce the yield of properly primed primer extension products, reducing the sensitivity of detection. The yield of primer extension products also can be adversely affected by template or primer degradation (e.g., by a nuclease activity of a polymerase). Mispriming and template or primer degradation can occur, e.g., when nucleic acid synthesis mixtures containing template, primers and polymerase are maintained at temperatures associated with manufacture, shipping, storage or bench top assembly of such mixtures.